Methods and pharmaceutical compositions for the treatment and prophylaxis of microbial infectious diseases and associated inflammatory disorders and for the treatment and prophylaxis of aging and associated diseases

ABSTRACT

A pharmaceutical composition and method are provided which may be used for the treatment and prevention of various infections, diseases, and conditions. Preferably, the composition and method may be used for preventing and treating of aging and aging associated disease and its use (or method). Preferably, the composition and method may be used for treating and prophylaxis of microbial infectious diseases and associated inflammatory disorders and its use (or method). The pharmaceutical composition according to the invention includes a therapeutic agent X or a pharmaceutically acceptable salt thereof; a therapeutic agent Y or a pharmaceutically acceptable salt thereof; and at least one pharmaceutically acceptable excipient, in which the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority to and the benefit of the filing dateof U.S. Provisional Application No. 63/124,905, filed on Dec. 14, 2020,entitled “METHODS AND COMPOSITIONS FOR THE TREATMENT AND PROPHYLAXIS OFMICROBIAL INFECTIOUS DISEASES AND ASSOCIATED INFLAMMATORY DISORDERS”,which is hereby incorporated by reference in its entirety. Thisapplication also claims priority to and the benefit of the filing dateof U.S. Provisional Application No. 63/139,337, filed on Jan. 20, 2021,entitled “METHODS AND COMPOSITIONS FOR THE TREATMENT AND PROPHYLAXIS OFAGING AND ASSOCIATED DISEASES”, which is hereby incorporated byreference in its entirety.

FIELD OF THE INVENTION

This patent specification relates to the field of treatment andprevention of diseases and infections. More specifically, this patentspecification relates to the use of small molecules to prevent or treataging and associated diseases and to the use of small molecules preventor treat microbial infection.

BACKGROUND

There are many diseases and infections which affect the humans and othermammals. This includes infectious diseases and associated inflammatorydisorders and also aging and associated diseases.

Microbial infectious disease is the combinatorial process of invasion ofinfectious agents into the organism, the replication of these agents andthe reaction of host tissue against these agents, includinginflammation, adaptive immune responses and sometimes a cytokine storm.Examples of microbes include viruses, bacteria, fungi, protozoans andparasites. Microbes replicate inside host cells and can produce toxinsthat cause disease. Microbial infections often show different degrees ofseverity in different infected persons. In mild cases, infections can bemanifested as fever, chills, fatigue, cough, diarrhea and othersymptoms, and severe cases can be manifested as dyspnea, increased heartrate, heart fibrillations, chest pain, acute lung injury (ALI), acuterespiratory distress syndrome (ARDS), cytokine release syndrome (CRS),thrombosis, tissue inflammation, tissue damage, acute organ failure,sepsis, and even death. It has been discussed that the severity ofmicrobial infections is heavily dependent on host responses. Hence, itwill be of great importance to target microbial infections by modulatingthe internal environment of the host, by modulating systems endogenousto the host.

As mentioned, microbial infection can be caused by viruses. Virusinfections can be caused by a DNA virus or an RNA virus such as membersof the Myoviridae, Podoviridae, Siphoviridae, Alloherpesviridae,Herpesviridae (including human herpes virus, and Varicella zostervirus), Malocoherpesviridae, Lipothrixviridae, Rudiviridae,Adenoviridae, Ampullaviridae, Ascoviridae, Asfarviridae (includingAfrican swine fever virus), Baculoviridae, Cicaudaviridae, Clavaviridae,Corticoviridae, Fuselloviridae, Globuloviridae, Guttaviridae,Hytrosaviridae, Iridoviridae, Maseilleviridae, Mimiviridae, Nudiviridae,Nimaviridae, Pandoraviridae, Papillomaviridae, Phycodnaviridae,Plasmaviridae, Polydnaviruses, Polyomaviridae (including Simian virus40, JC virus, BK virus), Poxviridae (including Cowpox and smallpox),Sphaerolipoviridae, Tectiviridae, Turriviridae, Dinodnavirus,Salterprovirus, Rhizidovirus, Coronaviridae, Picornaviridae,Caliciviridae, Flaviviridae, Togaviridae, Bornaviridae, Filoviridae,Paramyxoviridae, Pneumoviridae, Rhabdoviridae, Arenaviridae,Bunyaviridae, Orthomyxoviridae, or Deltavirus. A viral infection mayfurther be caused by a Coronavirus, Poliovirus, Rhinovirus, Hepatitis A,Hepatitis B virus, Norwalk virus, Yellow fever virus, West Nile virus,human immunodeficiency virus, Hepatitis C virus, Dengue fever virus,Zika virus, Rubella virus, Ross River virus, Sindbis virus, Chikungunyavirus, Borna disease virus, Ebola virus, Marburg virus, Measles virus,Mumps virus, Nipah virus, Hendra virus, Newcastle disease virus, Humanrespiratory syncytial virus, Rabies virus, Lassa virus, Hantavirus,Crimean-Congo hemorrhagic fever virus, Influenza, or Hepatitis D virus.

Severity of viral infection can be divided into mild, moderate, severe,and critically ill according to clinical manifestations. Currentanti-viral therapies include antibody cocktails or drugs targetingprocesses critical to the life cycles of the virus such as viral entry,viral uncoating, viral replication, viral synthesis, viral assembly, orviral release. These aforementioned strategies tend to target aparticular virus or particular class of virus, hence limiting theirapplication in the face of a different viral outbreak.

Ageing is associated with a progressive degeneration of the tissues,which has a negative impact on the structure and function of vitalorgans and is among the most important known risk factors for mostchronic diseases. Given the proportion of the world's populationaged >60 years doubles over the next four decades, the increasedincidences of chronic age-related diseases will place a huge burden onhealthcare resources.

Today, we are beginning to understand that aging is a ubiquitous complexphenomenon that results from environmental, stochastic, genetic, andepigenetic events in different cells and tissues and their interactionsthroughout life. A pervasive feature of aging tissues and age-relateddiseases is chronic inflammation. “Inflammaging” describes thelow-grade, chronic, systemic inflammation in aging, in the absence ofovert infection (“sterile” inflammation), and is a highly significantrisk factor for both morbidity and mortality in the elderly people.There is overwhelming epidemiological evidence that a state of mildinflammation, revealed by elevated levels of inflammatory biomarkers isassociated and predictive of many aging phenotypes—for example, changesin body composition, energy production and utilization, metabolichomeostasis, immune senescence, and neuronal health.

Targeting pathways that control age-related inflammation across multiplesystems may therefore be beneficial in old people or in treating agingrelated disorders or syndromes.

Therefore, a need exists for novel methods and compositions for thetreatment and prophylaxis of microbial infectious diseases andassociated inflammatory disorders. A need also exists for novel methodsand compositions for the treatment and prophylaxis of aging andassociated diseases.

BRIEF SUMMARY OF THE INVENTION

The present application provides pharmaceutical compositions and methodsof using a drug combination which may be used to treat and preventvarious aging related disorders and syndromes and which may also be usedto improve the host response, for prophylaxis and treatment ofinfectious diseases and their associated inflammatory disorders.

According to one aspect consistent with the principles of the invention,a pharmaceutical composition is provided which may be used forpreventing and treating of infectious disease. According to anotheraspect, the pharmaceutical composition may be used for preventing andtreating of aging and aging associated disease.

In some embodiments, the pharmaceutical composition may comprise atherapeutic agent X or a pharmaceutically acceptable salt thereof; atherapeutic agent Y or a pharmaceutically acceptable salt thereof; andat least one pharmaceutically acceptable excipient, in which thetherapeutic agent X is a non-steroidal anti-inflammatory drug and thetherapeutic agent Y is a fatty acid oxidation inhibitor. Thepharmaceutical composition according to the invention may be used in amethod of treating one or more conditions in a subject in need thereof,the conditions including the prevention and treatment of infectiousdisease and the prevention and treatment of aging associated diseasesand syndromes. A method of treating a condition in a subject in need maycomprise administering to the subject an effective amount of thepharmaceutical composition that comprises a therapeutic agent X or apharmaceutically acceptable salt thereof; a therapeutic agent Y or apharmaceutically acceptable salt thereof; and at least onepharmaceutically acceptable excipient, in which the therapeutic agent Xis a non-steroidal anti-inflammatory drug and the therapeutic agent Y isa fatty acid oxidation inhibitor.

In further embodiments, in the pharmaceutical composition, thetherapeutic agent X and the therapeutic agent Y are included in a singledosage form.

In further embodiments, in the pharmaceutical composition, thetherapeutic agent X and the therapeutic agent Y exist in separate dosageforms.

In further embodiments, the therapeutic agent X is selected from atleast one of the COX inhibitors, salicylates, ibuprofen, indomethacin,flurbiprofen, phenoxyibuprofen, naproxen, nabumetone, piroxicam,butazone, diclofenac, fenoprofen, ketoprofen, ketorolac,tetrachlorofenoic acid, sulindac and tometine. In some embodiments,agent X is a salicylate or a derivative thereof. In some embodiments,agent X is 2-acetyloxybenzoic acid.

In further embodiments, the therapeutic agent Y is selected from atleast one of the CPT inhibitors, carnitine biosynthesis inhibitors,3-ketoacyl-coenzyme A thiolase inhibitors, etomoxir, oxfenicine,perhexiline, mildronate, trimetazidine, ethoxylcarnitine, aminocarnitineor a phosphonyloxy derivative of carnitine. In some embodiments, agent Yis 1-[(2,3,4-trimethoxyphenyl)methyl]piperazine.

In some embodiments, the weight ratio of the therapeutic agent X and thetherapeutic agent Y of the pharmaceutical composition may be between 1:1to 10:1, including 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1.In further embodiments, the mass ratio of the therapeutic agent X andthe therapeutic agent Y may be between 1:1 to 10:1, including 1:1, 2:1,3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1.

In some embodiments, the pharmaceutical composition may be administeredin an oral dosage form. In further embodiments, the pharmaceuticalcomposition may be administered in injection form.

In some embodiments, the therapeutic agent X and the therapeutic agent Yare administered simultaneously. In some embodiments, the therapeuticagent X and the therapeutic agent Y are included in a single dosageform. In some embodiments, the single drug dosage form is an oral dosageform. In further embodiments, the single drug dosage form is aninjection form.

In some embodiments, the therapeutic agent X and the therapeutic agent Yare administered respectively. In some embodiments, the therapeuticagent X is administered prior to the therapeutic agent Y. In someembodiments, the therapeutic agent X is administered after thetherapeutic agent Y. In some embodiments, therapeutic agent X andtherapeutic agent Y are administered orally, respectively. In someembodiments, therapeutic agent X and therapeutic agent Y areadministered by injection, respectively.

BRIEF DESCRIPTION OF THE DRAWINGS

Some embodiments of the present invention are illustrated as an exampleand are not limited by the figures of the accompanying drawings, inwhich like references may indicate similar elements and in which:

FIG. 1A-FIG. 1A shows the scheme for microbial disease modeling and drugdosing.

FIG. 1B-FIG. 1B shows the drug combination X and Y prevents microbialLPS-induced death in mice.

FIG. 1C-FIG. 1C shows the drug combination X and Y ameliorates microbialLPS-induced lung inflammation in mice.

FIG. 2 -FIG. 2 shows that the drug combination X and Y decreasesexpression of the hepatitis C virus coreceptor CD36.

FIG. 3 -FIG. 3 shows that the drug combination X and Y decreasesexpression of the hepatitis C virus coreceptor CD81.

FIG. 4 -FIG. 4 shows that the drug combination X and Y decreasesexpression of the hepatitis A virus coreceptor HAVCR1.

FIG. 5 -FIG. 5 shows that the drug combination X and Y decreasesexpression of the hepatitis A virus coreceptor HAVCR2

FIG. 6 -FIG. 6 shows that the drug combination X and Y decreasesexpression of the coronavirus coreceptor ACE2.

FIG. 7 -FIG. 7 shows that the drug combination X and Y decreasesexpression of the coronavirus coreceptor DPP4

FIG. 8 -FIG. 8 shows that the drug combination X and Y decreasesexpression of the virus coreceptor integrin alpha(v).

FIG. 9 -FIG. 9 shows that the drug combination X and Y decreasesexpression of the virus coreceptor integrin beta1.

FIG. 10 -FIG. 10 shows that the drug combination X and Y decreasesexpression of the virus coreceptor integrin beta3.

FIG. 11 -FIG. 11 shows that the drug combination X and Y decreasesexpression of the virus coreceptor integrin beta5.

FIG. 12 -FIG. 12 shows that the drug combination X and Y decreasesexpression of the virus coreceptor integrin beta6.

FIG. 13A-FIG. 13A shows that the drug combination X and Y suppressessenescence biomarkers and reduces senescent cells in aged liver tissues.

FIG. 13B-FIG. 13B shows that the drug combination X and Y suppressessenescence biomarkers and reduces senescent cells in aged muscletissues.

FIG. 13C-FIG. 13C shows that the drug combination X and Y suppressessenescence biomarkers and reduces senescent cells in aged skin tissues.

FIG. 14 -FIG. 14 shows that the drug combination X and Y acceleratestissue repair in aged mice with chronic wounds.

FIG. 15 -FIG. 15 shows that the drug combination X and Y accelerateshair growth in aged mice with alopecia.

FIG. 16 -FIG. 16 shows that the drug combination X and Y amelioratestissue fibrosis in aged mice.

FIG. 17 -FIG. 17 shows that the drug combination X and Y amelioratesinflammaging and inflammation-induced death in mice.

FIG. 18A-FIG. 18A shows that the drug combination X and Y reversesinflammaging-associated and immunosenescence-associated gene signatures.

FIG. 18B-FIG. 18B shows that the drug combination X and Y reversesinflammaging-associated and immunosenescence-associated gene signatures.

FIG. 18C-FIG. 18C shows that the drug combination X and Y reversesinflammaging-associated and immunosenescence-associated gene signatures.

FIG. 19A-FIG. 19A shows that drug combination X and Y promotes oxidativemuscle growth.

FIG. 19B-FIG. 19B shows that drug combination X and Y promotes oxidativemuscle growth.

FIG. 19C-FIG. 19C shows that drug combination X and Y promotes oxidativemuscle growth.

FIG. 19D-FIG. 19D shows that drug combination X and Y promotes oxidativemuscle growth.

FIG. 20 -FIG. 20 shows that drug combination X and Y induces manymyokines/adipokines, as physical exercise does.

FIG. 21A-FIG. 21A shows that X and Y bound to mitochondrial proteinUQCRH directly according to thermal proteome profiling with TMTpro16plex kit and Q-Exactive HF-X mass spectrometry(Thermo Fisher).

FIG. 21B-FIG. 21B shows that X and Y bound to mitochondrial proteinNDUFS6 directly according to thermal proteome profiling with TMTpro16plex kit and Q-Exactive HF-X mass spectrometry (Thermo Fisher).

FIG. 21C-FIG. 21C shows that X and Y bound to mitochondrial proteinCOX7A2 directly according to thermal proteome profiling with TMTpro16plex kit and Q-Exactive HF-X mass spectrometry (Thermo Fisher).

FIG. 22 -FIG. 22 shows that X and Y synergistically decreased ATP levelswithin minutes.

FIG. 23 -FIG. 23 shows that X and Y synergistically decreased acety-CoAand protein acetylation.

FIG. 24 -FIG. 24 shows that X and Y synergistically increased Pink1 forParkin-driven mitophagy according to immunostaining with Abcam ab23707antibody.

FIG. 25A-FIG. 25A shows that X and Y synergistically increasedmitochondrial-targeted myoblast protein levels.

FIG. 25B-FIG. 25B shows that X and Y synergistically increasedmitochondrial-targeted myotube protein levels.

FIG. 26 -FIG. 26 shows that X and Y synergistically increasedmitochondrial DNA levels according to qRT-PCR for mitochondrial genomicNADH dehydrogenases ND1 and ND4 relative to nuclear genomic B2M.

FIG. 27 -FIG. 27 shows that X and Y synergistically increased theIRE1a-mediated unfolded protein response (UPR) according to RNAseq(Illumina HiSeq).

FIG. 28A-FIG. 28A shows that X and Y synergistically decreased themitochondrial reactive oxygen species (ROS).

FIG. 28B-FIG. 28B shows that X and Y synergistically decreased themitochondrial reactive oxygen species (ROS).

FIG. 29A-FIG. 29A shows that X and Y synergistically increased theactivity of a variety of cytokine pathways (FIG. 17 ), includingsemaphorins, indicating that the drug combination X and Y delays agingby stimulating the neuroendocrine system and mimicking calorierestriction.

FIG. 29B-FIG. 29B shows that X and Y synergistically increased theactivity of a variety of cytokine pathways (FIG. 17 ), including GDNF,indicating that the drug combination X and Y delays aging by stimulatingthe neuroendocrine system and mimicking calorie restriction.

FIG. 29C-FIG. 29C shows that X and Y synergistically increased theactivity of a variety of cytokine pathways (FIG. 17 ), including NGF,indicating that the drug combination X and Y delays aging by stimulatingthe neuroendocrine system and mimicking calorie restriction.

FIG. 30A-FIG. 30A shows that the drug combination X and Ysynergistically increased the expression of stem cell pathways accordingto RNAseq (Illumina Hiseq), such as Hippo, Notch-Hes-Hey, HIF1a, andintegrin-MAPK, indicating increased stemness and regenerative capacity.

FIG. 30B-FIG. 30B shows that the drug combination X and Ysynergistically increased the expression of stem cell pathways accordingto RNAseq (Illumina Hiseq), such as Hippo, Notch-Hes-Hey, HIF1a, andintegrin-MAPK, indicating increased stemness and regenerative capacity.

FIG. 30C-FIG. 30C shows that the drug combination X and Ysynergistically increased the expression of stem cell pathways accordingto RNAseq (Illumina Hiseq), such as Hippo, Notch-Hes-Hey, HIF1a, andintegrin-MAPK, indicating increased stemness and regenerative capacity.

FIG. 30D-FIG. 30D shows that the drug combination X and Ysynergistically increased the expression of stem cell pathways accordingto RNAseq (Illumina Hiseq), such as Hippo, Notch-Hes-Hey, HIF1a, andintegrin-MAPK, indicating increased stemness and regenerative capacity.

FIG. 30E-FIG. 30E shows that the drug combination X and Ysynergistically increased the expression of stem cell pathways accordingto RNAseq (Illumina Hiseq), such as Hippo, Notch-Hes-Hey, HIF1a, andintegrin-MAPK, indicating increased stemness and regenerative capacity.

FIG. 31 -FIG. 31 shows that after 2 rounds of transdermal treatment withX and Y, the cynomolgus monkeys' total body weight and waistcircumference (an indicator of aging-induced central adiposity) bothdecreased significantly by ˜10-20%, while the calf circumference (anindicator of skeletal muscle growth and regeneration) increased by ˜8%.

FIG. 32A-FIG. 32A shows that the cynomolgus monkeys' plasma levels ofthe neuroendocrine cytokine BDNF (R&D Systems ELISA) were alsosignificantly increased after treatment with the drug combination X andY, but not X alone or Y alone or the vehicle.

FIG. 32B-FIG. 32B shows that the cynomolgus monkeys' plasma levels ofthe neuroendocrine cytokine LIF (R&D Systems ELISA) were alsosignificantly increased after treatment with the drug combination X andY, but not X alone or Y alone or the vehicle.

FIG. 33A-FIG. 33A shows that no toxicities associated with drugcombination X and Y were observed according to clinical hematology orclinical blood chemistry (FIG. 21 ).

FIG. 33B-FIG. 33B shows that no toxicities associated with drugcombination X and Y were observed according to clinical hematology orclinical blood chemistry (FIG. 21 ).

FIG. 34 -FIG. 34 shows that X and Y promoted liver regeneration in1-year-old aged diabetic mice with livers with aging-associatedsteatohepatitis, relative to X alone or Y alone or the vehicle asobserved from increased hepatocyte proliferation indicated by Ki67(Abcam ab16667) immunostaining.

FIG. 35 -FIG. 35 shows that drug combination X and Y decreased β isletfibrosis in pancreas with aging-associated pancreatitis due to type 2diabetes, according to Masson's trichrome staining.

FIG. 36A-FIG. 36A shows that drug combination X and Y increased β isletcell growth and proliferation according to Ki67 (Abcam ab16667)immunostaining FIG. 36B-FIG. 36B shows that drug combination X and Yincreased β islet cell growth and proliferation according to Ki67 (Abcamab16667) immunostaining

FIG. 37A-FIG. 37A shows that drug combination X and Y increased β isletcell growth and proliferation according to qRT-PCR for Pax6, Mafa, andPdx1 using primers from Origene, thereby promoting pancreatic β isletregeneration.

FIG. 37B-FIG. 37B shows that drug combination X and Y increased β isletcell growth and proliferation according to qRT-PCR for Ins1 and Ins2using primers from Origene, thereby promoting pancreatic β isletregeneration.

FIG. 38 -FIG. 38 shows that drug combination X and Y promoted both agedhair regeneration and wound healing by 64 days.

FIG. 39 -FIG. 39 shows that, X and Y increased the protein levels ofmyosin heavy chain (MHC; Sigma-Aldrich-M4276 antibody) in the quadricepsmuscles of 1-year-old diabetic mice, thereby promoting skeletal muscleregeneration.

FIG. 40 -FIG. 40 shows that drug combination X and Y increased totalbody mass significantly, compared to the aged control animals withmuscle wasting and sarcopenia, indicating X and Y reversed sarcopenia.

FIG. 41 -FIG. 41 shows that daily oral gavage of the drug combination Xand Y, relative to X alone or Y alone or the vehicle over 3 weeks, ledto decreases in aging-associated lung fibrosis in 1 year-old aged micetreated with intratracheal bleomycin (3 mg/kg in 50 ul phosphatebuffered saline or PBS) to model chronic obstructive pulmonary disease(COPD) and idiopathic pulmonary fibrosis (IPF).

FIG. 42 -FIG. 42 shows that drug combination X and Y restored estruscycles in most of female rats (PCOS rat models), based on vaginal smearanalysis.

DETAILED DESCRIPTION OF THE INVENTION

The terminology used herein is for the purpose of describing particularembodiments only and is not intended to be limiting of the invention. Asused herein, the term “and/or” includes any and all combinations of oneor more of the associated listed items. As used herein, the singularforms “a,” “an,” and “the” are intended to include the plural forms aswell as the singular forms, unless the context clearly indicatesotherwise. It will be further understood that the terms “comprises”and/or “comprising,” when used in this specification, specify thepresence of stated features, steps, operations, elements, and/orcomponents, but do not preclude the presence or addition of one or moreother features, steps, operations, elements, components, and/or groupsthereof.

Unless otherwise defined, all terms (including technical and scientificterms) used herein have the same meaning as commonly understood by onehaving ordinary skill in the art to which this invention belongs. Itwill be further understood that terms, such as those defined in commonlyused dictionaries, should be interpreted as having a meaning that isconsistent with their meaning in the context of the relevant art and thepresent disclosure and will not be interpreted in an idealized or overlyformal sense unless expressly so defined herein.

The term “microbe” refers to tiny organisms that include virus,bacteria, fungi, protozoa, algae, amoebas, and slime molds and otherpotential disease-causing organisms.

The term “infection” refers to the invasion of an organism's bodytissues by disease-causing agents, their multiplication, and thereaction of host tissues to the infectious agents and the toxins theyproduce.

The term “aging-associated diseases” refers to diseases where incidencesof diseases increase with age and diseases that are degenerative.

The term “inflammaging” refers to the invasion of an organism's bodytissues by disease-causing agents, their multiplication, and thereaction of host tissues to the infectious agents and the toxins theyproduce.

The term “NSAID” or “nonsteroidal anti-inflammatory drugs” are usedherein interchangeably to refer to members of a drug class that inhibitsthe activity of cyclooxygenase enzymes (COX) to reduce pain, decreasefever and prevent drug clots. It is known in the art that not allinflammatory disorders can be treated with NSAIDs.

The terms “fatty acid oxidation” and “FAO” are used hereininterchangeably to refer to the biochemical process of breaking down afatty acid into acetyl-CoA units. In some embodiments, the FAO is in themitochondria of a cell. In some embodiments, the FAO is in theperoxisome of a cell. In some embodiments, the FAO rate-limiting step iscatalyzed by carnitine palmitoyltransferase (CPT).

As used herein, “treatment” or “treating” is an approach for obtainingbeneficial or desired results including clinical results. For purposesof this invention, beneficial or desired clinical results include, butare not limited to, one or more of the following: decreasing one or moresymptoms resulting from the disease, diminishing the extent of thedisease, stabilizing the disease or condition (e.g., preventing ordelaying the worsening of the disease or condition), preventing ordelaying the spread of the disease or condition, preventing or delayingthe occurrence or recurrence of the disease or condition, delaying orslowing the progression of the disease or condition, ameliorating thedisease state, providing a remission (whether partial or total) of thedisease or condition, decreasing the dose of one or more othermedications required to treat the disease or condition, delaying theprogression of the disease or condition, increasing the quality of life,and/or prolonging survival. Also encompassed by “treatment” is areduction of pathological consequence of the disease or condition. Themethods of the present application utilize any one or more of theseaspects of treatment.

As used herein, the term “prevention” and “prophylaxis” refers to ameasure for preventing or protecting against a disease, disorder orcondition from occurring in subjects that are at risk of disease but hasnot yet been diagnosed. Prevention (and effective preventive doses) orprophylaxis can be demonstrated in population studies, e.g., relative tothe untreated control subjects, an effective amount to prevent orprotect against a given disease or medical condition is an effective wayto reduce the incidence in the treated subjects.

The terms “individual,” “subject” and “patient” are used interchangeablyherein to describe a mammal, including humans. An individual includes,but is not limited to, human, bovine, ovine, porcine, equine, feline,canine, rodent, or primate. In some embodiments, the individual ishuman. In some embodiments, an individual suffers from a disease orcondition. In some embodiments, the individual is in need of treatment.

As is understood in the art, an “effective amount” refers to an amountof a composition sufficient to produce a desired therapeutic outcome.For therapeutic use, beneficial or desired results include, e.g.,decreasing one or more symptoms resulting from the disease or condition(biochemical, histologic and/or behavioral), including its complicationsand intermediate pathological phenotypes presented during development ofthe disease or condition, increasing the quality of life of thosesuffering from the disease or condition, decreasing the dose of othermedications required to treat the disease or condition, enhancing effectof another medication, delaying the progression of the disease orcondition, and/or prolonging survival of patients.

The term “pharmaceutically acceptable salt” means a salt which isacceptable for administration to a patient, such as a mammal (salts withcounterions having acceptable mammalian safety for a given dosageregime). Such salts can be derived from pharmaceutically acceptableinorganic or organic bases and from pharmaceutically acceptableinorganic or organic acids. “Pharmaceutically acceptable salt” refers topharmaceutically acceptable salts of a compound, which salts are derivedfrom a variety of organic and inorganic counter ions well known in theart and include, by way of example only, sodium, potassium, calcium,magnesium, ammonium, tetraalkylammonium, and the like; and when themolecule contains a basic functionality, salts of organic or inorganicacids, such as hydrochloride, hydrobromide, formate, tartrate, besylate,mesylate, acetate, maleate, oxalate, and the like.

The term “salt thereof” means a compound formed when a proton of an acidis replaced by a cation, such as a metal cation or an organic cation andthe like. Where applicable, the salt is a pharmaceutically acceptablesalt, although this is not required for salts of intermediate compoundsthat are not intended for administration to a patient. By way ofexample, salts of the present compounds include those wherein thecompound is protonated by an inorganic or organic acid to form a cation,with the conjugate base of the inorganic or organic acid as the anioniccomponent of the salt.

The terms “X and Y” or “X+Y” or “XY” are used interchangeably herein torefer to a combination comprising of both therapeutic agent X orpharmaceutically acceptable salts thereof and therapeutic agent Y orpharmaceutically acceptable salts thereof.

Both therapeutic agent X or pharmaceutically acceptable salts thereofand therapeutic agent Y can either be administered together oradministered one after another.

In describing the invention, it will be understood that a number oftechniques and steps are disclosed. Each of these has individual benefitand each can also be used in conjunction with one or more, or in somecases all, of the other disclosed techniques. Accordingly, for the sakeof clarity, this description will refrain from repeating every possiblecombination of the individual steps in an unnecessary fashion.Nevertheless, the specification and claims should be read with theunderstanding that such combinations are entirely within the scope ofthe invention and the claims.

As used in this application, the term “about” or “approximately” refersto a range of values within plus or minus 20% of the specified number.Additionally, as used in this application, the term “substantially”means that the actual value is within about 10% of the actual desiredvalue, particularly within about 5% of the actual desired value andespecially within about 1% of the actual desired value of any variable,element or limit set forth herein.

New methods and compositions for the treatment and prophylaxis of agingand associated diseases and for the treatment and prophylaxis ofmicrobial infectious diseases and associated inflammatory disorders arediscussed herein. In the following description, for purposes ofexplanation, numerous specific details are set forth in order to providea thorough understanding of the present invention. It will be evident,however, to one skilled in the art that the present invention may bepracticed without these specific details.

The present disclosure is to be considered as an exemplification of theinvention, and is not intended to limit the invention to the specificembodiments illustrated by the figures or description below.

The present invention will now be described by example and throughreferencing the appended figures representing preferred and alternativeembodiments. According to an embodiment of the present invention acomposition of matter is provided which may be used to treat variousdiseases, conditions, and infections. In some embodiments, thecomposition of matter may comprise a pharmaceutical composition having atherapeutic agent X or a pharmaceutically acceptable salt thereof; atherapeutic agent Y or a pharmaceutically acceptable salt thereof; andat least one pharmaceutically acceptable excipient, in which thetherapeutic agent X is a non-steroidal anti-inflammatory drug and thetherapeutic agent Y is a fatty acid oxidation inhibitor. In someembodiments, therapeutic agent X is a salicyclic acid or derivativethereof (e.g. salsalate). In some embodiments, therapeutic agent X is2-acetyloxybenzoic acid. In some embodiments, therapeutic agent Y is1-[(2,3,4-trimethoxyphenyl)methyl]piperazine. According to anotherembodiment of the present invention a method of treating one or moreconditions in a subject in need thereof, the conditions including theprevention and treatment of infectious disease and the prevention andtreatment of aging associated diseases and syndromes is provided. Amethod of treating a condition in a subject in need may compriseadministering to the subject an effective amount of the pharmaceuticalcomposition that comprises a therapeutic agent X or a pharmaceuticallyacceptable salt thereof; a therapeutic agent Y or a pharmaceuticallyacceptable salt thereof; and at least one pharmaceutically acceptableexcipient, in which the therapeutic agent X is a non-steroidalanti-inflammatory drug and the therapeutic agent Y is a fatty acidoxidation inhibitor.

The pharmaceutical composition and method comprising the administrationof the pharmaceutical composition according to the invention is able toeffectively treat or prevent microbial infections.

In some embodiments, the pharmaceutical composition comprisingtherapeutic agent X or a pharmaceutically acceptable salt thereof and atherapeutic agent Y or a pharmaceutically acceptable salt thereof mayeffectively prevent and or treat viral infections by modulating theimmune system. In some preferred embodiments, the pharmaceuticalcomposition comprising therapeutic agent X or a pharmaceuticallyacceptable salt thereof and a therapeutic agent Y or a pharmaceuticallyacceptable salt thereof may regulate immunomodulatory factors such asbut not limited to: ACVR1, ACVR1B, ACVR2A, ACVR2B, ACVRL1, AMH, AMHR2,BMP2, BMP7, BMPR1A, BMPR1B, BMPR2, CCL11, CCL15, CCL17, CCL19, CCL2,CCL20, CCL21, CCL22, CCL23, CCL24, CCL27, CCL28, CCL3, CCL4, CCL5, CCL7,CCL8, CCR1, CCR10, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CD27,CD40, CD40LG, CD70, CLCF1, CNTF, CNTFR, CRLF2, CSF1, CSF1R, CSF2,CSF2RA, CSF2RB, CSF3, CSF3R, CTF1, CX3CL1, CX3CR1, CXCL10, CXCL11,CXCL12, CXCL13, CXCL14, CXCL16, CXCL6, CXCL9, CXCR1, CXCR2, CXCR3,CXCR4, CXCR5, CXCR6, EDA, EDA2R, EDAR, EGF, EGFR, EPO, EPOR, FAS, FASLG,FLT1, FLT3, FLT3LG, FLT4, GDF5, GH1, GHR, HGF, IFNA1, IFNA10, IFNA17,IFNA2, IFNA4, IFNA5, IFNA6, IFNA7, IFNA8, IFNAR1, IFNAR2, IFNB1, IFNE,IFNG, IFNGR1, IFNGR2, IFNK, IFNL3, IFNLR1, IL10, IL10RA, IL10RB, IL11,IL11RA, IL12A, IL12B, IL12RB1, IL12RB2, IL13, IL13RA1, IL15, IL15RA,IL17A, IL17B, IL17RA, IL17RB, IL18, IL18R1, IL18RAP, IL19, ILIA, IL1B,IL1R1, IL1R2, IL1RAP, IL2, IL20, IL20RA, IL20RB, IL21, IL21R, IL22,IL22RA1, IL22RA2, IL23A, IL23R, IL24, IL25, IL2RA, IL2RB, IL2RG, IL3,IL4, IL4R, IL5, IL5RA, IL6, IL6R, IL6ST, IL7, IL7R, IL9, IL9R, INHBA,INHBB, INHBC, INHBE, KDR, KIT, KITLG, LEP, LEPR, LIF, LIFR, LTA, LTB,MET, MPL, NGFR, OSM, OSMR, PDGFA, PDGFB, PDGFC, PDGFRA, PDGFRB, PLEKHO2,PPBP, PRL, PRLR, RELT, TGFB1, TGFB2, TGFB3, TGFBR1, TGFBR2, TNFA,TNFRSF10B, TNFRSF10C, TNFRSF10D, TNFRSF11A, TNFR5F11B, TNFRSF12A,TNFRSF13B, TNFRSF13C, TNFRSF14, TNFRSF17, TNFRSF18, TNFRSF19, TNFRSF1A,TNFRSF1B, TNFRSF21, TNFRSF25, TNFRSF4, TNFRSF8, TNFRSF9, TNFSF10,TNFSF11, TNFSF12, TNFSF13, TNFSF13B, TNFSF14, TNFSF15, TNFSF18, TNFSF4,TNFSF8, TNFSF9, TPO, TSLP, VEGFA, VEGFB, VEGFC, VEGFD, XCL1, XCR1. Inanother aspect, the invention provides a method of treating a conditionin a subject in need by regulating one or more of the aboveimmunomodulatory factors, the method comprising administering to thesubject an effective amount of the pharmaceutical composition.

In some embodiments, the pharmaceutical composition comprisingtherapeutic agent X or a pharmaceutically acceptable salt thereof and atherapeutic agent Y or a pharmaceutically acceptable salt thereof mayeffectively prevent and or treat viral infections by modulatinginflammation. In some preferred embodiments, the pharmaceuticalcomposition comprising therapeutic agent X or a pharmaceuticallyacceptable salt thereof and a therapeutic agent Y or a pharmaceuticallyacceptable salt thereof may decrease the expression of factors such asbut not limited to: AKT1, AKT2, AKT3, BTK, CASP8, CCL3, CCL4, CCL5,CD14, CD180, CD40, CD80, CD86, CHUK, CISH, CTNNAL1, CUEDC2, CXCL10,CXCL11, CXCL8, CXCL9, CYLD, FADD, FBXW5, FOS, GM-CSF, IFNA1, IFNA10,IFNA13, IFNA14, IFNA16, IFNA17, IFNA2, IFNA21, IFNA4, IFNA5, IFNA6,IFNA7, IFNA8, IFNAR1, IFNAR2, IFNB1, IFNG, IKBKB, IKBKE, IKBKG, IL12A,IL12B, IL18, IL1B, IL6, IL8, IRAK1, IRAK2, IRAK3, IRAK4, IRF3, IRF5,IRF7, JUN, LBP, LY96, MAP2K1, MAP2K2, MAP2K3, MAP2K4, MAP2K6, MAP2K7,MAP3K7, MAP3K8, MAPK1, MAPK10, MAPK11, MAPK12, MAPK13, MAPK14, MAPK3,MAPK8, MAPK9, MBL2, MIR105-1, MIR6502, MIR718, MIR98, MIRLET7E,MIRLET7I, MLST8, MYD88, NFKB1, NFKB2, NFKBIA, OTUD5, PELI1, PELI2,PELI3, PIK3CA, PIK3CB, PIK3CD, PIK3CG, PIK3R1, PIK3R2, PIK3R3, PIK3R5,PLK1, PROBE, PTPN6, RAC1, RBCK1, RELA, RIPK1, RNF216, RNF31, RNF41,SARM1, SFTPA2, SFTPD, SIGIRR, SMAD6, SOCS1, SPP1, SQSTM1, STAT1, SYK,TAB1, TAB2, TABS, TBK1, TICAM1, TICAM2, TIFA, TIRAP, TLR1, TLR2, TLR3,TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TMED7, TNFA, TNFAIP3, TOLLIP, TRAF3,TRAF6, TRAFD1, TREM1, USP7, ZMYND11. In another aspect, the inventionprovides a method of treating a condition in a subject in need bydecreasing the expression of one or more of the above factors, themethod comprising administering to the subject an effective amount ofthe pharmaceutical composition.

In some embodiments, the pharmaceutical composition comprisingtherapeutic agent X or a pharmaceutically acceptable salt thereof and atherapeutic agent Y or a pharmaceutically acceptable salt thereof mayeffectively prevent and or treat viral infections by decreasing a viralreceptor. In some preferred embodiments, the pharmaceutical compositioncomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may decrease the expression of factors such as but not limitedto: ACE2, CD4, CD36, CD81, CLDN1, CXCR4, DPP4, HAVCR1, HAVCR2, ITGAV,ITGB1, ITGB3, ITGB5, ITGB6, MAG, SLC1A4, SLC1A5, SLC7A1, SLC20A1,SLC20A2. In another aspect, the invention provides a method of treatinga condition in a subject in need by decreasing the expression of one ormore of the above factors, the method comprising administering to thesubject an effective amount of the pharmaceutical composition.

In some embodiments, the pharmaceutical composition comprisingtherapeutic agent X or a pharmaceutically acceptable salt thereof and atherapeutic agent Y or a pharmaceutically acceptable salt thereof mayeffectively prevent and or treat microbial infections caused by a virus,a bacterium, a fungus, a protozoan, or a parasite. In furtherembodiments, a method of effectively preventing and/or treatingmicrobial infections caused by a virus, a bacterium, a fungus, aprotozoan, or a parasite, may comprise administering to the subject aneffective amount of the pharmaceutical composition.

In embodiments where the infection is viral, it may be caused by a DNAvirus. In some specific embodiments, the DNA virus may include, but isnot necessarily limited to members of the Adenoviridae, Papovaviridae,Parvoviridae, Anelloviridae, Pleolipoviridae, Myoviridae, Podoviridae,Siphoviridae, Alloherpesviridae, Herpesviridae (including human herpesvirus, and Varicella zoster virus), Malocoherpesviridae,Lipothrixviridae, Rudiviridae, Adenoviridae, Ampullaviridae,Ascoviridae, Asfarviridae (including African swine fever virus),Baculoviridae, Cicaudaviridae, Clavaviridae, Corticoviridae,Fuselloviridae, Globuloviridae, Guttaviridae, Hytrosaviridae,Iridoviridae, Maseilleviridae, Mimiviridae, Nudiviridae, Nimaviridae,Pandoraviridae, Papillomaviridae, Phycodnaviridae, Plasmaviridae,Polydnaviruses, Polyomaviridae (including Simian virus 40, JC virus, BKvirus), Poxviridae (including Cowpox and smallpox), Sphaerolipoviridae,Tectiviridae, Turriviridae, Dinodnavirus, Salterprovirus, orRhizidovirus.

A viral infection may also be caused by a double-stranded RNA virus, apositive sense RNA virus, a negative sense RNA virus, a retrovirus, or acombination thereof. A viral infection may further be caused by aCoronaviridae virus, a Picornaviridae virus, a Reoviridae virus, aCaliciviridae virus, a Flaviviridae virus, a Togaviridae virus, aBornaviridae virus, a Filoviridae virus, a Paramyxoviridae virus, aPneumoviridae virus, a Rhabdoviridae virus, an Arenaviridae, aBunyaviridae virus, an Astroviridae virus an Orthomyxoviridae virus, anArteriviridae virus, a Hepeviridae virus, a Retroviridae virus, aCaulimoviridae virus, a Hepadnaviridae virus or a Deltavirus. A viralinfection may further be caused by a Coronavirus, Poliovirus,Rhinovirus, Hepatitis A, Norwalk virus, Yellow fever virus, West Nilevirus, Hepatitis C virus, Dengue fever virus, Zika virus, Rubella virus,Ross River virus, Sindbis virus, Chikungunya virus, Borna disease virus,Ebola virus, Marburg virus, Measles virus, Mumps virus, Nipah virus,Hendra virus, Newcastle disease virus, Human respiratory syncytialvirus, Rabies virus, Lassa virus, Hantavirus, Crimean-Congo hemorrhagicfever virus, Influenza, or Hepatitis D virus. In some preferredembodiments, the viral infection is caused by a coronavirus. Examples ofa coronavirus includes but is not limited to Alphacoronavirus 1, Humancoronavirus 229E, Human coronavirus NL63, Miniopterus bat coronavirus 1,Miniopterus bat coronavirus HKU8, Porcine epidemic diarrhea virus,Rhinolophus bat coronavirus HKU2, Scotophilus bat coronavirus 512,Betacoronavirus 1 (Bovine Coronavirus, Human coronavirus OC43), Hedgehogcoronavirus 1, Human coronavirus HKU1, Middle East respiratorysyndrome-related coronavirus, Murine coronavirus, Pipistrellus batcoronavirus HKU5, Rousettus bat coronavirus HKU9, Severe acuterespiratory syndrome-related coronavirus (SARS-CoV, SARS-CoV-2),Tylonycteris bat coronavirus HKU4, Avian coronavirus, Beluga whalecoronavirus SW1, Bulbul coronavirus HKU11 and Porcine coronavirus HKU15.In some preferred embodiments, the viral infection is caused bySARS-CoV2.

In other embodiments, the infection may be bacterial in nature. Thebacterium causing the bacterial infection may include, but is notnecessarily limited to, Acinetobacter species, Actinobacillus species,Actinomycetes species, an Actinomyces species, Aerococcus species anAeromonas species, an Anaplasma species, an Alcaligenes species, aBacillus species, a Bacteroides species, a Bartonella species, aBifidobacterium species, a Bordetella species, a Borrelia species, aBrucella species, a Burkholderia species, a Campylobacter species, aCapnocytophaga species, a Chlamydia species, a Citrobacter species, aCoxiella species, a Corynbacterium species, a Clostridium species, anEikenella species, an Enterobacter species, an Escherichia species, anEnterococcus species, an Ehrlichia species, an Epidermophyton species,an Erysipelothrix species, a Eubacterium species, a Francisella species,a Fusobacterium species, a Gardnerella species, a Gemella species, aHaemophilus species, a Helicobacter species, a Kingella species, aKlebsiella species, a Lactobacillus species, a Lactococcus species, aListeria species, a Leptospira species, a Legionella species, aLeptospira species, Leuconostoc species, a Mannheimia species, aMicrosporum species, a Micrococcus species, a Moraxella species, aMorganell species, a Mobiluncus species, a Micrococcus species,Mycobacterium species, a Mycoplasm species, a Nocardia species, aNeisseria species, a Pasteurelaa species, a Pediococcus species, aPeptostreptococcus species, a Pityrosporum species, a Plesiomonasspecies, a Prevotella species, a Porphyromonas species, a Proteusspecies, a Providencia species, a Pseudomonas species, aPropionibacteriums species, a Rhodococcus species, a Rickettsia species,a Rhodococcus species, a Serratia species, a Stenotrophomonas species, aSalmonella species, a Serratia species, a Shigella species, aStaphylococcus species, a Streptococcus species, a Spirillum species, aStreptobacillus species, a Treponema species, a Tropheryma species, aTrichophyton species, an Ureaplasma species, a Veillonella species, aVibrio species, a Yersinia species, a Xanthomonas species, orcombination thereof.

In other embodiments, the infection may be fungal, and may be caused byfungi such as, but not necessarily limited to, Aspergillus, Blastomyces,Candidiasis, Coccidiodomycosis, Cryptococcus neoformans, Cryptococcusgatti, sp. Histoplasma sp. (such as Histoplasma capsulatum),Pneumocystis sp. (such as Pneumocystis jirovecii), Stachybotrys (such asStachybotrys chartarum), Mucroymcosis, Sporothrix, fungal eye infectionsringworm, Exserohilum, Cladosporium, Geotrichum, Saccharomyces, aHansenula species, a Candida species, a Kluyveromyces species, aDebaryomyces species, a Pichia species, a Penicillium species, aCladosporium species, a Byssochlamys species or a combination thereof.

In other embodiments, the infection may be caused by a protozoan, suchas Euglenozoa, a Heterolobosea, a Diplomonadida, an Amoebozoa, aBlastocystic, an Apicomplexa, or a combination thereof.

In other embodiments, the infection may be caused by a parasite, suchas, but not necessarily limited to, Trypanosoma cruzi (Chagas disease),T. brucei gambiense, T. brucei rhodesiense, Leishmania braziliensis, L.infantum, L. mexicana, L. major, L. tropica, L. donovani, Naegleriafowleri, Giardia intestinalis (G. lamblia, G. duodenalis), canthamoebacastellanii, Balamuthia madrillaris, Entamoeba histolytica, Blastocystichominis, Babesia microti, Cryptosporidium parvum, Cyclosporacayetanensis, Plasmodium falciparum, P. vivax, P. ovale, P. malariae,and Toxoplasma gondii, or a combination thereof.

In some embodiments, the pharmaceutical composition and methodcomprising the administration of the pharmaceutical compositionaccording to the invention is able to treat and/or prevent an associatedinflammatory disorder that is related to cytokine and immunedysfunction, such as, but not limited to CRS, ARDS, ALI, AIDS, asthma,chronic peptic ulcer, tuberculosis, rheumatoid arthritis, periodontitis,ulcerative colitis, Crohn's disease, sinusitis, encephalitis, myelitis,meningitis, arachnoiditis, neuritis, dacryoadenitis, scleritis,episcleritis, keratitis, retinitis, chorioretinitis, blepharitis,conjunctivitis, uveitis, otitis externa, otitis media, Labyrinthitis,mastoiditis, carditis, endocarditis, myocarditis, pericarditis,vasculitis, arteritis, phlebitis, capillaritis, sinusitis, rhinitis,pharyngitis, laryngitis, tracheitis, bronchitis, bronchiolitis,pneumonitis, pleuritis, mediastinitis, stomatitis, gingivitis,gingivostomatitis, glossitis, tonsillitis, sialadenitis, parotitis,cheilitis, pulpitis, gnathitis, esophagitis, gastritis, gastroenteritis,enteritis, colitis, enterocolitis, duodenitis, ileitis, caecitis,appendicitis, proctitis, hepatitis, ascending cholangitis,cholecystitis, pancreatitis, peritonitis, dermatitis, folliculitis,cellulitis, hidradenitis, arthritis, dermatomyositis, myositis,synovitis, tenosynovitis, bursitis, enthesitis, fasciitis, capsulitis,epicondylitis, tendinitis, panniculitis, osteochondritis, osteitis,osteomyelitis, spondylitis, periostitis, chondritis, nephritis,glomerulonephritis, pyelonephritis, ureteritis, cystitis, urethritis,oophoritis, salpingitis, endometritis, parametritis, cervicitis,vaginitis, vulvitis, mastitis, orchitis, epididymitis, prostatitis,seminal vesiculitis, balanitis, posthitis, balanoposthitis,chorioamnionitis, funisitis, omphalitis, insulitis, hypophysitis,thyroiditis, parathyroiditis, adrenalitis, lymphangitis orlymphadenitis.

In some embodiments, the pharmaceutical composition and methodcomprising the administration of the pharmaceutical compositionaccording to the invention is able to treat and/or prevent aging-relateddiseases, the pharmaceutical composition comprising therapeutic agent Xor a pharmaceutically acceptable salt thereof and a therapeutic agent Yor a pharmaceutically acceptable salt thereof.

In some embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and/or treat aging associated diseasesby promoting regeneration.

In some embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat aging associated diseasesby reversing fibrosis.

In some embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat aging associated diseasesby reversing senescence. The effect of therapeutic agent X andtherapeutic agent Y is able to be evaluated by testing the therapeuticagents against relevant cell and tissue models, some of which aredetailed in the examples listed. Alternatively, the effect oftherapeutic agent X and therapeutic agent Y is able to be evaluated bytesting the therapeutic agents against relevant animal models, some ofwhich are described in the examples.

In some embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat aging associated diseasesby improving insulin sensitivity or preventing or treating insulinresistance. In some preferred embodiments, the pharmaceuticalcomposition comprising therapeutic agent X or a pharmaceuticallyacceptable salt thereof and a therapeutic agent Y or a pharmaceuticallyacceptable salt thereof may by improving insulin sensitivity, therebyimprove glucose tolerance.

In some embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat aging and aging associateddiseases by modulating inflammation. In some embodiments, thepharmaceutical composition comprising therapeutic agent X or apharmaceutically acceptable salt thereof and a therapeutic agent Y or apharmaceutically acceptable salt thereof may effectively prevent and ortreat aging and aging associated diseases by preventing or treating orreducing chronic inflammation or inflammaging. In some preferredembodiments, the pharmaceutical composition and method comprisingtherapeutic agent X or a pharmaceutically acceptable salt thereof and atherapeutic agent Y or a pharmaceutically acceptable salt thereof maydecrease the expression of factors such as but not limited to: AKT1,AKT2, AKT3, BTK, CASP8, CCL3, CCL4, CCL5, CD14, CD180, CD40, CD80, CD86,CHUK, CISH, CTNNAL1, CUEDC2, CXCL10, CXCL11, CXCL8, CXCL9, CYLD, FADD,FBXW5, FOS, GM-CSF, IFNA1, IFNA10, IFNA13, IFNA14, IFNA16, IFNA17,IFNA2, IFNA21, IFNA4, IFNA5, IFNA6, IFNA7, IFNA8, IFNAR1, IFNAR2, IFNB1,IFNG, IKBKB, IKBKE, IKBKG, IL12A, IL12B, IL18, IL1B, IL6, IL8, IRAK1,IRAK2, IRAK3, IRAK4, IRF3, IRF5, IRF7, JUN, LBP, LY96, MAP2K1, MAP2K2,MAP2K3, MAP2K4, MAP2K6, MAP2K7, MAP3K7, MAP3K8, MAPK1, MAPK10, MAPK11,MAPK12, MAPK13, MAPK14, MAPK3, MAPK8, MAPK9, MBL2, MIR105-1, MIR6502,MIR718, MIR98, MIRLET7E, MIRLET7I, MLST8, MYD88, NFKB1, NFKB2, NFKBIA,OTUD5, PELI1, PELI2, PELI3, PIK3CA, PIK3CB, PIK3CD, PIK3CG, PIK3R1,PIK3R2, PIK3R3, PIK3R5, PLK1, PROBE, PTPN6, RAC1, RBCK1, RELA, RIPK1,RNF216, RNF31, RNF41, SARM1, SFTPA2, SFTPD, SIGIRR, SMAD6, SOCS1, SPP1,SQSTM1, STAT1, SYK, TAB1, TAB2, TABS, TBK1, TICAM1, TICAM2, TIFA, TIRAP,TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TMED7, TNFA,TNFAIP3, TOLLIP, TRAF3, TRAF6, TRAFD1, TREM1, USP7, ZMYND11.

In some embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat aging associated diseasessuch as Alzheimer's Disease, Parkinson's Disease, dementiaatherosclerosis, cardiovascular disease, type II Diabetes, cancer,arthritis, rheumatoid arthritis, periodontitis, cataracts, osteoporosis,fibrosis, cirrhosis, idiopathic pulmonary fibrosis, cardiac fibrosis,uterine fibrosis, scarring, arthrofibrosis, chronic kidney disease,Crohn's disease, keloids, myelofibrosis, retroperitoneal fibrosis,scleroderma, sclerosis, chronic wounds (such as diabetic foot ulcer),chronic dermal fibrosis, cutaneous fibrosis, skin aging, nonalcoholicsteatohepatitis (NASH), hair loss, tissue atrophy, menopause, primaryovarian insufficiency, endometrial hyperplasia, adenomyosis andsarcopenia.

In some preferred embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat fibrosis. It is widelyaccepted in the state of the art that fibrosis, inflammation-inducedfibrosis, inflammation and inflammaging can be modeled in animals forexample by administering bleomycin or lipopolysaccharide to animalmodels (e.g., mice or rats).

In some preferred embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat pulmonary fibrosis.Non-limiting examples of pulmonary fibrosis include but are not limitedto Chronic obstructive pulmonary disease (COPD), Idiopathic pulmonaryfibrosis (IPF), Interstitial Lung Disease (ILD).

In some preferred embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat hair loss or alopecia.Non-limiting examples of hair loss include but are not limited toalopecia areata, alopecia totalis and alopecia universalis andandrogenic alopecia.

In some preferred embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat cirrhosis.

In some preferred embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat NASH.

In some preferred embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat type II diabetes.

In some preferred embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat chronic wounds.

In some preferred embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat aging-associated diseasesspecific to women such as but not limited to menopause, primary ovarianinsufficiency, endometrial hyperplasia, adenomyosis, polycystic ovarysyndrome.

In some preferred embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and/or treat sarcopenia.

In some preferred embodiments, the pharmaceutical composition and methodcomprising therapeutic agent X or a pharmaceutically acceptable saltthereof and a therapeutic agent Y or a pharmaceutically acceptable saltthereof may effectively prevent and or treat sarcopenia. Sarcopenia is acondition characterized by loss of skeletal muscle mass and function andoccurs in high prevalence among the aged, especially aged men. Increasein muscle mass may therefore be seen as a way to ameliorate or treatsuch a condition.

In some embodiments, the present invention may comprise a pharmaceuticalcomposition having a therapeutic agent X or a pharmaceuticallyacceptable salt thereof; a therapeutic agent Y or a pharmaceuticallyacceptable salt thereof; and at least one pharmaceutically acceptableexcipient, in which the therapeutic agent X is a non-steroidalanti-inflammatory drug and the therapeutic agent Y is a fatty acidoxidation inhibitor. In further embodiments, the present invention maycomprise a method of treating a condition in a subject in need thereof,the method comprising administering to the subject an effective amountof a pharmaceutical composition having a therapeutic agent X or apharmaceutically acceptable salt thereof; a therapeutic agent Y or apharmaceutically acceptable salt thereof; and at least onepharmaceutically acceptable excipient, in which the therapeutic agent Xis a non-steroidal anti-inflammatory drug and the therapeutic agent Y isa fatty acid oxidation inhibitor.

In some embodiments, in the pharmaceutical composition, the therapeuticagent X or a pharmaceutically acceptable salt thereof and thetherapeutic agent Y or a pharmaceutically acceptable salt thereof areincluded in a single dosage form.

In further embodiments, in the pharmaceutical composition, thetherapeutic agent X or a pharmaceutically acceptable salt thereof andthe therapeutic agent Y or a pharmaceutically acceptable salt exist inseparate dosage forms.

In some embodiments, the therapeutic agent X is selected from at leastone of the COX inhibitors, salicylates, ibuprofen, phenoxyibuprofen,naproxen, diclofenac, celecoxib, mefenamic acid, etoricoxib,indomethacin, ketorolac, tetrachlorofenoic acid, sulindac and tometine.In some embodiments, therapeutic agent X is a salicylate or derivativethereof. In some embodiments, therapeutic agent X is 2-acetyloxybenzoicacid. In some embodiments, the therapeutic agent Y is selected from atleast one of the CPT inhibitors, carnitine biosynthesis inhibitors,3-ketoacyl-coenzyme A thiolase inhibitors, etomoxir, oxfenicine,perhexiline, mildronate, 1-(2,3,4-Trimethoxybenzyl) piperazinedihydrochloride, trimetazidine, ethoxylcarnitine, aminocarnitine or aphosphonyloxy derivative of carnitine. In some preferred embodiments,therapeutic agent Y is 1-(2,3,4-Trimethoxybenzyl) piperazinedihydrochloride.

In some embodiments, the mass ratio of the therapeutic agent X and thetherapeutic agent Y is between 1:1 to 10:1, including 1:1, 2:1, 3:1,4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1. In some preferred embodiments,the mass ratio of the therapeutic agents is 5:1 or 6:1 or 7:1.

In some embodiments, the amount of therapeutic agent X can rangeanywhere from 1 mg/kg to 100 mg/kg or more per day and the amount oftherapeutic agent Y can range anywhere from 0.25 mg/kg to 100 mg/kg ormore per day. The exact amount to be used varies according the exactdisease indication and the physiology of the individual. In someembodiments, therapeutic agent X and therapeutic agent Y can beadministered once in n days, where n ranges from 1 to 14. In someembodiments, therapeutic agent X and therapeutic agent Y can beadministered once a day or multiple times a day. In some embodiments,therapeutic agent X and therapeutic agent Y can be administered everyday, every two days, every three days, every four days, every five days,every six days, weekly, etc.

In some embodiments, the therapeutic agent X and the therapeutic agent Yare administered simultaneously. In some embodiments, the therapeuticagent X and the therapeutic agent Y are included in a single dosageform. In some embodiments, the possible routes of administration oftherapeutic agent X and therapeutic agent Y in a single dosage forminclude but is not limited to: oral, sublingual, buccal, nasal,inhalation, intratracheal, intravenous, intraarterial, intracoronary,intrathecal, intramuscular, intraperitoneal, intramyocardial,trans-endocardial, trans-epicardial, subcutaneous, transdermal, vaginal,rectal, or otic. In some preferred embodiments, therapeutic agent X andtherapeutic agent Y in a single dose form are administered orally orintravenously or subcutaneously.

In some embodiments, the therapeutic agent X and the therapeutic agent Yare administered respectively. In some embodiments, the therapeuticagent X is administered prior to the therapeutic agent Y. In someembodiments, the therapeutic agent X is administered after thetherapeutic agent Y. In some embodiments, the possible routes ofadministration of therapeutic agent X and therapeutic agent Y inseparate dosage forms include but is not limited to: oral, sublingual,buccal, nasal, inhalation, intratracheal, intravenous, intraarterial,intracoronary, intrathecal, intramuscular, intraperitoneal,intramyocardial, trans-endocardial, trans-epicardial, subcutaneous,transdermal, vaginal, rectal, or otic. In some embodiments, therapeuticagent X and therapeutic agent Y in separate dosage forms areadministered orally or intravenously.

In some embodiments, therapeutic agent X and therapeutic agent Y areadministered orally, respectively. In some embodiments, therapeuticagent X and therapeutic agent Y are administered by injection,respectively.

Pharmaceutical compositions for parenteral injection include aqueous ornon-aqueous solutions, dispersions, suspensions, or emulsions,pharmaceutically acceptable sterile or non-sterile, and powders forreconstitution in sterile injectable solutions or dispersions.

In some embodiments, and for a more effective distribution, thecompounds of the invention may be embedded in extended controlledrelease or directed administration systems, such as polymeric matrices,liposomes and microspheres.

In preferred embodiments, the pharmaceutical composition of the presentinvention may be a solid composition for oral administration, andspecific examples thereof include tablets, granules, fine granules,capsules, powders, and pills. The solid oral composition of the presentinvention may have an excipient, a binder, a lubricant and the likeadded thereto, in addition to the pharmaceutically acceptable salt oftherapeutic agent X or therapeutic agent Y or combinations thereof, andmay be formulated into the respective forms. Examples of excipients thatmay be used include lactose, corn starch, crystalline cellulose,sucrose, glucose, mannitol, sorbitol, and calcium carbonate. Examples ofthe binder include hydroxypropyl cellulose, hydroxyethyl cellulose,hypromellose, hydroxyethylethyl cellulose, hydroxyethylmethyl cellulose,polyvinylpyrrolidone, and polyvinyl alcohol. Examples of the lubricantinclude magnesium stearate, stearic acid, palmitic acid, calciumstearate, and talc. Such formulations are described in detail in anumber of sources which are well known and readily available to thoseskilled in the art. For example, Remington's The Science and Practice ofPharmacy, edited by Allen, Loyd V., Jr, 22nd edition, describe themaking of formulations which may be used in connection with the subjectinvention.

The examples below are intended to be purely exemplary of the presentapplication and should therefore not be considered to limit theinvention in any way. The following examples and detailed descriptionsare offered by way of illustration and not by way of limitation.

Example 1: LPS-Induced Inflammation

Lipopolysaccharide (LPS) is a common antigen expressed by manypathogenic microbes, with severe effects on the host response that arenot treatable with non-steroidal anti-inflammatory drugs alone. Inparticular, intraperitoneal treatment with LPS is a reliable model ofpathogen-induced CRS, that also recapitulates many features of humanARDS, including enhanced production of pro-inflammatory factors, highlevels of immune infiltration, and loss of endothelial and vascularintegrity in the lung.

Adult 6-8 week-old DIO mice were fed with high fat diet (Jackson Labs)for 12 weeks before any experiments were initiated. DIO mice received 5mg/kg of LPS (Sigma) injected intraperitoneally. Mice on a normal chowdiet served as controls. Mice injected with PBS vehicle served ascontrols for drug treatment. Mice were killed at 6 hours afterstimulation with LPS (FIG. 1A). Lungs were frozen in liquid nitrogen formolecular assays, or fixed in formalin for histologic examination.Histology results showed severe pulmonary congestion, alveolarthickening, hyaline membrane formations, and inflammatory infiltrationin lung tissues from all LPS-treated mice, but these changes were morepronounced in DIO mice. Lungs from DIO mice exhibited more severe signsof ALI compared with normal mice, as shown by the increased lung proteincontent, cell number in bronchoalveolar lavage fluid (BALF), and lacticacid dehydrogenase (LDH) content in BALF. These results show thatdiet-induced obese (DIO) mice are more susceptible to LPS-induced lunginjury (FIG. 1B).

To test effects on mortality and survival, LPS was injectedintraperitoneally at a dose of 30 mg/kg. The mortality of mice wasrecorded every 12 hours for 3 days after the LPS injection (n=12).LPS-treated DIO mice started to die by 16 hours, and ˜80% of the micewere dead by 40 hours. In contrast, normal mice treated with LPS livedlonger, and only ˜40% died by 40 hours. These results showed thatdiet-induced obese (DIO) mice are more susceptible to death fromLPS-induced inflammation (FIG. 1B).

Further assessments of the infiltration of inflammatory cells into lungsshowed that lung myeloperoxidase (MPO) activity and the number ofneutrophils in BALF were higher in DIO mice compared to normal mice. Thelevels of TNF-a in BALF and lung tissue were also much higher in DIOmice than normal mice treated with LPS. These results show that obesityexacerbates LPS-induced lung inflammation (FIG. 1C).

To test whether prophylactic treatment with the drugs X and Y may blockALI pathogenesis, we performed a rescue experiment using intraperitonealinjections of X and Y. In this experiment, and in examples listedhenceforth, X is 2-acetoxybenzoic acid (CAS no: 50-78-2) and Y is1-[(2,3,4-trimethoxyphenyl)methyl]piperazine, (CAS no: 5011-34-7).Histologic analysis showed that the lung structure of DIO miceprophylactically treated with the drug combination XY 2 weeks before LPSinjection, was significantly improved in comparison to DIO mice treatedwith X alone, Y alone, or PBS vehicle, 2 weeks before LPS injection.These results show that the drug combination XY ameliorates ALI in DIOmice (FIG. 1B).

Survival analysis further showed that the survival rate of DIO miceprophylactically treated with the drug combination XY 2 weeks before LPSinjection, was significantly improved in comparison to DIO mice treatedwith X alone, Y alone, or PBS vehicle, 2 weeks before LPS injection.These results show that the drug combination XY prevents ALI-induceddeath in DIO mice (FIG. 1B).

Moreover, the edema formation protein, cell number, LDH content andneutrophil counts in BALF, and MPO activity of DIO mice prophylacticallytreated with the drug combination XY 2 weeks before LPS injection, wereall significantly attenuated in comparison to DIO mice treated with Xalone, Y alone, or PBS vehicle, 2 weeks before LPS injection. Theseresults show that the drug combination XY ameliorates LPS-induced lunginflammation (FIG. 1C).

Example 2: Virus Coreceptors

To test if the drug treatment might be effective in reducing viralcoreceptor expression, with potential efficacy in reducing virusinfection, we performed RNAseq profiling of various organs after 3 daysof XY treatment. Briefly, total RNA was purified by ethanolprecipitation and sent for RNAseq. Paired-end libraries were constructedand sequenced using a Nova-PE150 platform (Illumina). FIGS. 2 and 3 showthat the drug combination XY decreases expression of the hepatitis Cvirus coreceptors CD36 and CD81. FIGS. 4 and 5 show that the drugcombination XY decreases expression of the hepatitis A virus coreceptorsHAVCR1 and HAVCR2. FIG. 6 shows that the drug combination XY decreasesexpression of the coreceptor for SARS-CoV and SARS-CoV-2, ACE2, as wellas the coreceptor for MERS coronavirus DPP4. FIGS. 7-12 show that thedrug combination XY decreases expression of integrin alpha(v), integrinbeta1, integrin beta3, integrin beta5, integrin beta6, which arecoreceptors for adenoviruses, parvoviruses and cytomegaloviruses. Theseresults show that the drug combination XY may mitigate the spread ofviral infectious disease.

Example 3: Senescent Cells in Aged Mammalian Tissues

Mammalian cells that exhibit senescence are marked by positive stainingfor senescence-associated-β-galactosidase (SA-β-gal), and proteinexpression of p16INK4A and senescence-associated heterochromatin markerssuch as H3K9me3. To test if the drugs X and Y may reduce senescence inthe tissues of aged mice, we intraperitoneally injected 2-year-old agedmice (n=10/group) with 20 mg/kg of X, or 4 mg/kg of Y, or both 20 mg/kgof X and 4 mg/kg of Y, relative to the DMSO vehicle control, on a dailybasis for 30 days. Staining of liver, muscle and skin tissue sectionsfor SA-β-gal activity was performed using the Senescence β-GalactosidaseStaining Kit (Cell Signaling Technology) according to manufacturer'sprotocols. Protein expression of p16INK4A and H3K9me3 were evaluated bydensitometric quantification of Westem blots of the liver, muscle andskin tissue samples. The results revealed that only the drug combinationX and Y significantly decreased all three biomarkers of senescence inaged liver, muscle and skin tissues (P<0.01), but not X alone or Yalone, relative to the DMSO vehicle control (FIGS. 13A-13C). Thissuggests that the drug combination X and Y is able to decrease senescentcells and/or reverse senescence.

Example 4: Cutaneous Wound Healing and Tissue Repair During Aging

Mammals manifest reduced rates of tissue repair with aging. One animalmodel for this phenomenon is the delayed cutaneous wound healing in ageddb/db mice, compared to young mice. Chronic wounds were generated indb/db mice by performing full thickness 6 mm diameter excision wounds onthe dorsal skin of 9-month-old mice. We used a well-described murinefull-thickness excisional wound model (Loh et al., 2009). Briefly, a 6mm disposable biopsy punch (Delasco) was used to make two circular fullthickness wounds on the dorsal back skin of mice. Silicon wound splints(Grace BioLabs) were sutured with 4-0 Nylon to prevent skin contracture.Wounds were dressed with a sterile occlusive dressing and monitoreddaily. Borders were monitored by frequently application of permanentmarker. Photos were taken at various time points throughout the durationof the experiment. The wounds remained open 20 days after wounding, andsometimes for over 3 months, as expected of chronic wounds. To test ifthe drugs X and Y can treat the chronic wounds of aged mice, weintraperitoneally injected aged mice (n=10/group) 20 days afterwounding, with 20 mg/kg of X, or 4 mg/kg of Y, or both 20 mg/kg of X and4 mg/kg of Y, relative to the DMSO vehicle control, on a daily basis for30 days.

Quantification of the dorsal wound areas revealed that only the drugcombination X and Y significantly accelerated tissue repair in aged micewith chronic wounds (P<0.01), but not X alone or Y alone, relative tothe DMSO vehicle control (FIG. 14 ).

Example 5: Hair Loss and Hair Regrowth During Aging

Humans manifest hair loss or alopecia with aging, most obviously in menexhibiting androgenetic alopecia or senile alopecia. Inflammation andaging, or inflammaging, are risk factors for alopecia. An animal modelfor this phenomenon is the delayed hair regeneration in ob/ob mice withinflammaging, after depilation. After dorsal hair removal from8-week-old mice using depilatory cream (day 0), 3 wild-type mice and 3inflammaging mice were regularly monitored until the depilated area wasmostly covered with hair. Regenerated hair could be seen as dark hair onpinkish-white shaved skin. The hair regeneration area was quantifiedusing the NIH ImageJ software program (NTIS, Springfield, VA). When thehair cycle reaches the second telogen phase by 8 weeks age in wild-typemice, depilation induces a rapid transition to anagen by the 1st weekafter depilation and complete hair regeneration by the 4th week afterdepilation in wild-type mice. In contrast, the first signs of anagen wasonly observed at the 6th week after depilation in inflammaging mice. Totest if the drugs X and Y can ameliorate the hair regeneration defect ininflammaging mice, we intraperitoneally injected inflammaging mice afterdepilation (n=10/group), with 20 mg/kg of X, or 4 mg/kg of Y, or both 20mg/kg of X and 4 mg/kg of Y, relative to the DMSO vehicle control, on adaily basis for 30 days. Quantification of the dorsal skin area withhair regeneration revealed that only the drug combination X and Ysignificantly accelerated hair regrowth in inflammaging mice afterdepilation (P<0.01), but not X alone or Y alone, relative to the DMSOvehicle control (FIG. 15 ).

Example 6: Tissue Fibrosis and Tissue Degeneration During Aging

Aging mammals also display a reduced ability to resolve fibrosis,leading to tissue scarring and irreparable organ damage, e.g., inidiopathic pulmonary fibrosis (IPF), liver fibrosis and cardiacfibrosis. An animal model for IPF is bleomycin-induced lung injury inmice. Young and aged mice were anesthesized with intraperitonealinjection of ketamine (100 mg/kg) and xylazine (10 mg/kg), thenadministered with intra-tracheal bleomycin (1.25 U/kg) to induce lunginjury, or saline (50 μL total volume). Fibrosis was quantified bymeasuring the fibrotic areas in lung tissue sections with Massontrichrome staining. An animal model for both liver fibrosis and cardiacfibrosis is the inflammaging mouse model, i.e., ob/ob mice fed on a highfat, high sugar, high cholesterol diet for 90 days (Research Labs). Totest if the drugs X and Y can ameliorate the lung fibrosis inbleomycin-injured mice, and the liver and cardiac fibrosis ininflammaging mice, we intraperitoneally injected bleomycin-treated mice(n=10/group) and inflammaging mice (n=10/group) with 20 mg/kg of X, or 4mg/kg of Y, or both 20 mg/kg of X and 4 mg/kg of Y, relative to the DMSOvehicle control, on a daily basis for 30 days. Quantification of thefibrosis by Masson trichrome staining revealed that only the drugcombination X and Y significantly reversed fibrosis in inflammaging mice(P<0.01), but not X alone or Y alone, relative to the DMSO vehiclecontrol (FIG. 16 ).

Example 7: Inflammation and Death

Aging and especially inflammaging are also associated with increasedrisk of death from septic shock or pro-inflammatory cytokine releasesyndrome. This is partly due to inflammaging-induced immunosenescence,which leads to dysfunctional immune responses that cause multi-organfailure and death. An animal model for this phenomenon is LPSendotoxin-induced septic shock in aged mice. For lethality studies, LPS(Sigma Aldrich) was injected intraperitoneally at a dose of 30 mg/kg.The mortality of mice was recorded every 12 hours for 3 days after theLPS challenge in each group. To test if the drugs X and Y canprophylactically ameliorate the lethal effects of LPS-induced sepsis, weintraperitoneally injected 2-year-old aged mice (n=10/group) with 20mg/kg of X, or 4 mg/kg of Y, or both 20 mg/kg of X and 4 mg/kg of Y,relative to the DMSO vehicle control, on a daily basis for 30 days,before subjecting them to the LPS challenge. Quantification of thesurvival rate over time revealed that only the drug combination X and Ysignificantly improved survival after the LPS challenge (P<0.01), butnot X alone or Y alone, relative to the DMSO vehicle control (FIG. 17 ).

Example 8: Immunosenescence-Associated Gene Signatures

Aging and especially chronic inflammaging leads to immunosenescence,i.e., the decrease in naïve T cells and dysfunctional skewing of immunecell subsets with aging. This causes an aberrant spike in the Th17- andTh1-driven pro-inflammatory responses when immunologically challenged,at the expense of the Treg- and Th2-driven anti-inflammatory response,which could lead to a lethal cytokine release syndrome and/or sepsis. Totest how the drug combination X and Y affects the immune response, weisolated peripheral blood mononuclear cells (PBMCs) from mice treatedwith 20 mg/kg of X and 4 mg/kg of Y, relative to the DMSO vehiclecontrol, on a daily basis for 30 days, and extracted their RNA withTrizol (Invitrogen) for RNAseq (Illumina). After Gene Set EnrichmentAnalysis (GSEA; Broad Institute), our results showed that the drugcombination X and Y suppressed the Th1 pro-inflammatory signature, theTh1-associated/pro-Th17 cytokine IL2 signature, the Th1/Th17-associatedcytokine IFNG signature, and the pan-activated Th-associated cytokinesIL3/IL5/GM-CSF signature (FIGS. 18A and 18B). In contrast, X and Ypromoted the Th2/Treg-associated anti-inflammatory cytokine signaturesof IL4, IL10 and IL13 (FIG. 18C).

Example 9: Mimicry of Exercise to Suppress Inflammaging

Physical muscular exercise is known to ameliorate the effects of aging,including chronic inflammaging and immunosenescence. To test how thedrug combination X and Y affects the effects of physical exercise,cryosections of quadriceps muscles in inflammaging mice wereimmunostained for type I slow-twitch myofibers (FIG. 19A), and examinedwith fluorescence microscopy (Carl Zeiss). The results showed that thedrug combination X and Y more than doubled the area of type I myofibers(P<0.01), indicating that the drug combination X and Y mimicked andenhanced the effects of endurance exercise (FIG. 19B). Moreover,quantification of the cross-sectional areas of the myofibers indicatedthat the drug combination X and Y increased the proportion of myofiberswith areas above the threshold of 100 pixels (P<0.01), suggesting thatthe drug combination X and Y induced muscle hypertrophy like physicalexercise (FIG. 19C). Finally, grip strength was measured as the averageof 6 measurements of the maximal peak strength generated on a gripstrength meter (Bioseb), normalized to their body weight. The resultsshowed that the drug combination X and Y significantly increased thegrip strength of the inflammaging mice (P<0.01), indicating the drugcombination X and Y increases limb muscle strength like physicalexercise (FIG. 19D).

Example 10: Mimicry of Exercise Induces Secretory Factors to SuppressInflammaging

Physical muscular exercise is known to ameliorate the effects of aging,including chronic inflammaging and immunosenescence, by promoting thesecretion of pro-metabolism and immunomodulatory cytokines. To test howthe drug combination X and Y affects the effects of physical exercisevia such secretory factors, the muscles of inflammaging mice wereprofiled with RNAseq and the serum of inflammaging mice were profiledwith ELISA (R&D). The results showed that a variety of secretoryfactors, including myokines, were significantly increased by the drugcombination X and Y (FIG. 20 ), including Angptl8, Angptl3, Angptl4,Adiponectin, BDNF, IGF1, IL6, Irisin and LIF (P<0.01), indicating thatthe drug combination X and Y affects aging and inflammaging, at least inpart, by mimicking the effects of physical exercise and inducing manysecretory factors including myokines.

Example 11: Mimicry of Calorie Restriction Induces Anti-Aging GenePathways

Calorie restriction is known to ameliorate the effects of aging, byaltering cellular bioenergetic stress and promoting mitochondrialactivity, the unfolded protein response (UPR), the secretion ofbeneficial cytokines, and an increase in stem cell regeneration. To testhow the drug combination X and Y mimics the effects of calorierestriction (CR), we profiled the metabolism of primary human skeletalmuscle cells at passage 15 treated with X (100 mg/L) and Y (2 mg/L),compared to X alone (100 mg/L) or Y alone (2 mg/L). The results showedthat X and Y bound to mitochondrial proteins UQCRH, NDUFS6, and COX7A2directly according to thermal proteome profiling with TMTpro 16plex kitand Q-Exactive HF-X mass spectrometry (Thermo Fisher; FIGS. 21A-21C),synergistically decreased ATP levels within minutes (FIG. 22 ),synergistically decreased acetyl-CoA and protein acetylation (FIG. 23 ),synergistically increased Pink1 for Parkin-driven mitophagy according toimmunostaining with the Abcam ab23707 antibody (FIG. 24 ),synergistically increased mitochondrial-targeted protein levels (FIGS.25A and 25B), synergistically increased mitochondrial DNA levelsaccording to qRT-PCR for mitochondrial genomic ND1 and ND4 relative tonuclear genomic B2M (FIG. 26 ), synergistically increased theIRE1a-mediated unfolded protein response (UPR) according to RNAseq(Illumina HiSeq) and gene set enrichment analysis (GSEA) (FIG. 27 ), andsynergistically decreased the mitochondrial reactive oxygen species(ROS; FIGS. 28A and 28B). X and Y synergistically increased the activityof a variety of cytokine pathways according to gene set enrichmentanalysis (GSEA) (FIGS. 29A-29C), including NGF, GDNF, and semaphorins,indicating that the drug combination X and Y delays aging by stimulatingthe neuroendocrine system and mimicking calorie restriction. Finally,the drug combination X and Y synergistically increased the expression ofstem cell pathways according to RNAseq (Illumina Hiseq) and gene setenrichment analysis (GSEA), such as Hippo, Notch-Hes-Hey, HIF1a, andintegrin-MAPK (FIGS. 30A-30E), indicating increased stemness andregenerative capacity. None of these beneficial anti-aging effects wereobserved with either X alone or Y alone or the vehicle.

Example 12: Induction of Anti-Aging, Pro-Regenerative Responses In Vivo

Application of the drug combination X and Y in cynomolgus monkeysincreased neuroendocrine cytokines like calorie restriction, therebyincreasing anti-aging-associated muscle regeneration and decreasingaging-associated central adiposity. By dissolving X and Y in DURO-TAK87-2677 PSA (pressure-sensitive adhesive) solution, and coating it on arelease film (Scotchpak™ 1022), followed by bake-drying and laminatingwith a backing film (Scotchpak™ 1109), we constructed a drug-in-adhesivepatch for controlled transdermal release of the drugs X and Y every 7days. After 2 rounds of transdermal drug treatment, we found that thecynomolgus monkeys' total body weight and waist circumference (anindicator of aging-induced central adiposity) both decreasedsignificantly by ˜10-20%, while the calf circumference (an indicator ofskeletal muscle growth and regeneration) increased by ˜8% (FIG. 31 ).Importantly, their plasma levels of the neuroendocrine cytokines BDNFand LIF (R&D Systems ELISA) were also significantly increased aftertreatment with the drug combination X and Y, but not X alone or Y aloneor the vehicle (FIGS. 32A and 32B). No toxicities were observedaccording to clinical hematology or clinical blood chemistry (FIGS. 33Aand 33B).

Daily oral gavage of the drug combination X (20 mg/kg) and Y (4 mg/kg),relative to X alone (20 mg/kg) or Y alone (4 mg/kg) or the vehicle (DMSOin phosphate buffered saline or PBS) over 6 weeks, led to increases inanti-aging-associated tissue regeneration in 1 year-old aged diabeticmice. In their livers with aging-associated steatohepatitis, X and Ypromoted liver regeneration, as observed from increased hepatocyteproliferation indicated by Ki67 (Abcam ab16667) immunostaining (FIG. 34). In their pancreas with aging-associated pancreatitis due to type 2diabetes, X and Y decreased β islet fibrosis according to Masson'strichrome staining (FIG. 35 ) and increased β islet cell growth andproliferation according to Ki67 (Abcam ab16667) immunostaining (FIGS.36A and 36B) and qRT-PCR for Pax6, Mafa, Pdx1 and Ins using primers fromOrigene (FIGS. 37A and 37B), thereby promoting pancreatic β isletregeneration. In their skin, after shaving and wounding with dorsal skinpunches (10 mm diameter), X and Y promoted both aged hair regenerationand wound healing by 64 days (FIG. 38 ). In their quadriceps muscles, Xand Y increased the protein levels of myosin heavy chain (MHC;Sigma-Aldrich-M4276 antibody) (FIG. 39 ), thereby promoting skeletalmuscle regeneration. Moreover, total body mass increased significantly,compared to the aged control animals with muscle wasting and sarcopenia,indicating X and Y reversed sarcopenia (FIG. 40 ). None of thesebeneficial anti-aging effects were observed with either X alone or Yalone or the vehicle control.

Daily oral gavage of the drug combination X and Y, relative to X aloneor Y alone or the vehicle over 3 weeks, led to decreases inaging-associated lung fibrosis in 1 year-old aged mice treated withintratracheal bleomycin (3 mg/kg in 50 ul phosphate buffered saline orPBS) to model chronic obstructive pulmonary disease (COPD) andidiopathic pulmonary fibrosis (IPF). Masson trichrome staining andImageJ (NIH) quantification of the % fibrotic area revealed that lungsof mice treated with X and Y had significantly lower levels of fibrosis(FIG. 41 ). None of these beneficial anti-aging effects were observedwith either X alone or Y alone or the vehicle control.

Daily oral gavage of the drug combination X and Y, relative to X aloneor Y alone or the vehicle over 6 weeks, led to restoration of estruscycles in adult female rats treated with subcutaneousdehydroepiandrosterone (DHEA; 60 mg/kg body weight) and fed with a highfat-high sugar-high cholesterol diet ad libitum to model prematureovarian aging in polycystic ovary syndrome (PCOS). Vaginal smearanalysis revealed that most of the female rats treated with X and Y hadrestored estrus cycles (FIG. 42 ). None of these beneficial anti-agingeffects were observed with either X alone or Y alone or the vehiclecontrol, where all of the rats were frozen in either the estrus orproestrus stages.

Although the present invention has been illustrated and described hereinwith reference to preferred embodiments and specific examples thereof,it will be readily apparent to those of ordinary skill in the art thatother embodiments and examples may perform similar functions and/orachieve like results. All such equivalent embodiments and examples arewithin the spirit and scope of the present invention, are contemplatedthereby, and are intended to be covered by the following claims.

1) A composition of matter for use in preventing or treating a microbialinfection that comprises a combination of therapeutic agent X andacceptable salts thereof and therapeutic agent Y and acceptable saltsthereof, wherein the therapeutic agent X is a non-steroidalanti-inflammatory drug and the therapeutic agent Y is a fatty acidoxidation inhibitor. 2) A composition of matter for use in preventing ortreating inflammatory disorders associated with microbial infection thatcomprises a combination of therapeutic agent X and acceptable saltsthereof and therapeutic agent Y and acceptable salts thereof, whereinthe therapeutic agent X is a non-steroidal anti-inflammatory drug andthe therapeutic agent Y is a fatty acid oxidation inhibitor. 3) Thecomposition of matter of claim 1 for use in preventing or treatingmicrobial infection and associated inflammatory disorders by modulatinghost responses. 4) The composition of matter of claim 1 for use inpreventing or treating microbial infection and associated inflammatorydisorders by modulating host immune system. 5) The composition of matterof claim 1 for use in preventing or treating microbial infection andassociated inflammatory disorders by modulating host inflammation. 6)The composition of matter of claim 1 for use in preventing or treatingmicrobial infection and associated inflammatory disorders by decreasingviral receptors. 7) The composition of matter of claim 1, for use inpreventing or treating inflammatory disorders selected from thefollowing group: CRS, ARDS, ALI, AIDS, asthma, chronic peptic ulcer,tuberculosis, rheumatoid arthritis, periodontitis, ulcerative colitis,Crohn's disease, sinusitis, encephalitis, myelitis, meningitis,arachnoiditis, neuritis, dacryoadenitis, scleritis, episcleritis,keratitis, retinitis, chorioretinitis, blepharitis, conjunctivitis,uveitis, otitis externa, otitis media, Labyrinthitis, mastoiditis,carditis, endocarditis, myocarditis, pericarditis, vasculitis,arteritis, phlebitis, capillaritis, sinusitis, rhinitis, pharyngitis,laryngitis, tracheitis, bronchitis, bronchiolitis, pneumonitis,pleuritis, mediastinitis, stomatitis, gingivitis, gingivostomatitis,glossitis, tonsillitis, sialadenitis, parotitis, cheilitis, pulpitis,gnathitis, esophagitis, gastritis, gastroenteritis, enteritis, colitis,enterocolitis, duodenitis, Ileitis, caecitis, appendicitis, proctitis,hepatitis, ascending cholangitis, cholecystitis, pancreatitis,peritonitis, dermatitis, folliculitis, cellulitis, hidradenitis,arthritis, dermatomyositis, myositis, synovitis, tenosynovitis,bursitis, enthesitis, fasciitis, capsulitis, epicondylitis, tendinitis,panniculitis, osteochondritis, osteitis, osteomyelitis, spondylitis,periostitis, chondritis, nephritis, glomerulonephritis, pyelonephritis,ureteritis, cystitis, urethritis, oophoritis, salpingitis, endometritis,parametritis, cervicitis, vaginitis, vulvitis, mastitis, orchitis,epididymitis, prostatitis, seminal vesiculitis, balanitis, posthitis,balanoposthitis, chorioamnionitis, funisitis, omphalitis, insulitis,hypophysitis, thyroiditis, parathyroiditis, adrenalitis, lymphangitis orlymphadenitis. 8) The composition of matter of claim 1 for use inpreventing or treating microbial infection caused by bacteria, viruses,fungi, parasites, or protozoa. 9) The composition of matter of claim 1for use in preventing or treating microbial infection caused by a virus.10) The composition of matter of claim 1 for use in preventing ortreating microbial infection caused by a member of the Adenoviridae,Papovaviridae, Parvoviridae, Anelloviridae, Pleolipoviridae, Myoviridae,Podoviridae, Siphoviridae, Alloherpesviridae, Herpesviridae (includinghuman herpes virus, and Varicella zoster virus), Malocoherpesviridae,Lipothrixviridae, Rudiviridae, Adenoviridae, Ampullaviridae,Ascoviridae, Asfarviridae (including African swine fever virus),Baculoviridae, Cicaudaviridae, Clavaviridae, Corticoviridae,Fuselloviridae, Globuloviridae, Guttaviridae, Hytrosaviridae,Iridoviridae, Maseilleviridae, Mimiviridae, Nudiviridae, Nimaviridae,Pandoraviridae, Papillomaviridae, Phycodnaviridae, Plasmaviridae,Polydnaviruses, Polyomaviridae (including Simian virus 40, JC virus, BKvirus), Poxviridae (including Cowpox and smallpox), Sphaerolipoviridae,Tectiviridae, Turriviridae, Dinodnavirus, Salterprovirus, orRhizidovirus, Coronaviridae, Picornaviridae, Reoviridae, Caliciviridae,Flaviviridae, Togaviridae, Bornaviridae, Filoviridae, Paramyxoviridae,Pneumoviridae, Rhabdoviridae, Arenaviridae, Bunyaviridae, Astroviridae,Orthomyxoviridae, Arteriviridae, Hepeviridae, Retroviridae,Caulimoviridae, Hepadnaviridae or a Deltavirus, or combinations thereof.11) The composition of matter of claim 1 where the viral infection iscaused by a Coronavirus, Cytomegalovirus, Poliovirus, Rhinovirus,Hepatitis A, Hepatitis B virus, Norwalk virus, Yellow fever virus, WestNile virus, Hepatitis C virus, Dengue fever virus, Zika virus, Rubellavirus, Ross River virus, Sindbis virus, Chikungunya virus, Borna diseasevirus, Ebola virus, Marburg virus, Measles virus, Mumps virus, Nipahvirus, Hendra virus, Newcastle disease virus, Human respiratorysyncytial virus, Human immunodeficiency virus, Rabies virus, Lassavirus, Hantavirus, Crimean-Congo hemorrhagic fever virus, Influenza, orHepatitis D virus. 12) The composition of matter of claim 1 whereintherapeutic agent X or acceptable salts thereof and therapeutic agent Yor acceptable salts thereof are included in a single dosage form. 13)The composition of matter of claim 1 wherein therapeutic agent X oracceptable salts thereof and therapeutic agent Y or acceptable saltsthereof are included in separate dosage forms. 14) The composition ofmatter of claim 1 wherein the mass ratio of the therapeutic agent X andthe therapeutic agent Y is 1:1 to 10:1, including 2:1, 3:1, 4:1, 5:1,6:1, 7:1, 8:1, 9:1, or 10:1. 15) The composition of matter of claim 1wherein routes of administration to a subject include oral, sublingual,buccal, nasal, inhalation, intratracheal, intravenous, intraarterial,intracoronary, intrathecal, intramuscular, intraperitoneal,intramyocardial, trans-endocardial, trans-epicardial, subcutaneous,transdermal, vaginal, rectal, or otic. 16) The composition of matter ofclaim 1, wherein therapeutic agent X is selected from at least one ofCOX inhibitors, salicylates, ibuprofen, phenoxyibuprofen, naproxen,diclofenac, celecoxib, mefenamic acid, etoricoxib, indomethacin,ketorolac, tetrachlorofenoic acid, sulindac and tometine. 17) Thecomposition of matter of claim 1 wherein therapeutic agent Y is selectedfrom at least one of CPT inhibitors, carnitine biosynthesis inhibitors,3-ketoacyl-coenzyme A thiolase inhibitors, etomoxir, oxfenicine,perhexiline, mildronate, trimetazidine, ethoxylcarnitine, aminocarnitineor a phosphonyloxy derivative of carnitine. 18) The composition ofmatter for preventing or treating aging or aging-related diseases thatcomprises a combination of therapeutic agent X and acceptable saltsthereof and therapeutic agent Y and acceptable salts thereof, whereinthe therapeutic agent X is a non-steroidal anti-inflammatory drug andthe therapeutic agent Y is a fatty acid oxidation inhibitor. 19)(canceled) 20) (canceled) 21) (canceled) 22) (canceled) 23) (canceled)24) (canceled) 25) (canceled) 26) (canceled) 27) (canceled) 28)(canceled) 29) (canceled) 30) (canceled)